{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T03:01:23Z","timestamp":1787022083314,"version":"build-2736575974"},"reference-count":26,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2021,5,10]],"date-time":"2021-05-10T00:00:00Z","timestamp":1620604800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"DOI":"10.13039\/100000069","name":"National Institute of Arthritis and Musculoskeletal and Skin Diseases","doi-asserted-by":"publisher","award":["AR049794"],"award-info":[{"award-number":["AR049794"]}],"id":[{"id":"10.13039\/100000069","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["GM125503"],"award-info":[{"award-number":["GM125503"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000069","name":"National Institute of Arthritis and Musculoskeletal and Skin Diseases","doi-asserted-by":"publisher","award":["AR074993"],"award-info":[{"award-number":["AR074993"]}],"id":[{"id":"10.13039\/100000069","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000738","name":"U.S. Department of Veterans Affairs","doi-asserted-by":"publisher","award":["BX000294"],"award-info":[{"award-number":["BX000294"]}],"id":[{"id":"10.13039\/100000738","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>\n                    The cytokine RANKL is essential for osteoclast formation during physiological and pathological bone resorption. RANKL also contributes to lymphocyte production, development of lymph nodes and mammary glands, as well as other biological activities. Transcriptional control of the\n                    <jats:italic>Tnfsf11<\/jats:italic>\n                    gene, which encodes RANKL, is complex and involves distant regulatory regions. Nevertheless, cell culture studies suggest that an enhancer region near the transcription start site is involved in the control of\n                    <jats:italic>Tnfsf11<\/jats:italic>\n                    expression by hormones such as 1,25-(OH)\n                    <jats:sub>2<\/jats:sub>\n                    vitamin D\n                    <jats:sub>3<\/jats:sub>\n                    and parathyroid hormone, as well as the sympathetic nervous system. To address the significance of this region in vivo, we deleted the sequence between -510 to -1413 bp, relative to\n                    <jats:italic>Tnfsf11<\/jats:italic>\n                    exon 1, from mice using CRISPR-based gene editing. MicroCT analysis of the femur and fourth lumbar vertebra of enhancer knockout mice showed no differences in bone mass compared to wild type littermates at 5 weeks and 6 months of age, suggesting no changes in osteoclast formation. RNA extracted from the tibia, fifth lumbar vertebra, thymus, and spleen at 6 months of age also showed no reduction in\n                    <jats:italic>Tnfsf11<\/jats:italic>\n                    mRNA abundance between these groups. However, maximal stimulation of\n                    <jats:italic>Tnfsf11<\/jats:italic>\n                    mRNA abundance in cultured stromal cells by PTH was reduced approximately 40% by enhancer deletion, while stimulation by 1,25-(OH)\n                    <jats:sub>2<\/jats:sub>\n                    vitamin D\n                    <jats:sub>3<\/jats:sub>\n                    was unaffected. The abundance of B and T lymphocytes in the bone marrow did not differ between genotypes. These results demonstrate that the region between -510 and -1413 does not contribute to\n                    <jats:italic>Tnfsf11<\/jats:italic>\n                    expression, osteoclast support, or lymphocyte production in mice under normal physiological conditions but may be involved in situations of elevated parathyroid hormone.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0250974","type":"journal-article","created":{"date-parts":[[2021,5,10]],"date-time":"2021-05-10T14:16:56Z","timestamp":1620656216000},"page":"e0250974","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":7,"title":["Deletion of a putative promoter-proximal Tnfsf11 regulatory region in mice does not alter bone mass or Tnfsf11 expression in vivo"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7797-2754","authenticated-orcid":true,"given":"Ryan S.","family":"MacLeod","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark B.","family":"Meyer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinhu","family":"Xiong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keisha M.","family":"Cawley","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melda","family":"Onal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nancy A.","family":"Benkusky","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeff D.","family":"Thostenson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. Wesley","family":"Pike","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5286-6821","authenticated-orcid":true,"given":"Charles A.","family":"O\u2019Brien","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,5,10]]},"reference":[{"key":"pone.0250974.ref001","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/s41232-019-0111-3","article-title":"RANKL biology: bone metabolism, the immune system, and beyond","volume":"40","author":"T Ono","year":"2020","journal-title":"Inflammation and regeneration"},{"issue":"2","key":"pone.0250974.ref002","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/S0092-8674(00)81569-X","article-title":"Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation","volume":"93","author":"DL Lacey","year":"1998","journal-title":"Cell"},{"issue":"6717","key":"pone.0250974.ref003","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1038\/16852","article-title":"OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis","volume":"397","author":"YY Kong","year":"1999","journal-title":"Nature"},{"issue":"9","key":"pone.0250974.ref004","doi-asserted-by":"crossref","first-page":"5738","DOI":"10.4049\/jimmunol.0901563","article-title":"RANKL is necessary and sufficient to initiate development of antigen-sampling M cells in the intestinal epithelium","volume":"183","author":"KA Knoop","year":"2009","journal-title":"J Immunol"},{"issue":"4","key":"pone.0250974.ref005","doi-asserted-by":"crossref","first-page":"911","DOI":"10.1016\/j.bone.2009.08.050","article-title":"Control of RANKL gene expression","volume":"46","author":"CA O\u2019Brien","year":"2010","journal-title":"Bone"},{"issue":"27","key":"pone.0250974.ref006","doi-asserted-by":"crossref","first-page":"19301","DOI":"10.1074\/jbc.274.27.19301","article-title":"STAT3 activation in stromal osteoblastic cells is required for induction of the receptor activator of NF-kappa B ligand and stimulation of osteoclastogenesis by gp130-utilizing cytokines or interleukin-1 but not 1,25-dihydroxyvitamin D-3 or parathyroid hormone","volume":"274","author":"CA O\u2019Brien","year":"1999","journal-title":"Journal of Biological Chemistry"},{"issue":"17","key":"pone.0250974.ref007","doi-asserted-by":"crossref","first-page":"6453","DOI":"10.1128\/MCB.00356-06","article-title":"Parathyroid Hormone Controls Receptor Activator of NF-{kappa}B Ligand Gene Expression via a Distant Transcriptional Enhancer","volume":"26","author":"Q Fu","year":"2006","journal-title":"Mol Cell Biol"},{"issue":"17","key":"pone.0250974.ref008","doi-asserted-by":"crossref","first-page":"6469","DOI":"10.1128\/MCB.00353-06","article-title":"Activation of Receptor Activator of NF-{kappa}B Ligand Gene Expression by 1,25-Dihydroxyvitamin D3 Is Mediated through Multiple Long-Range Enhancers","volume":"26","author":"S Kim","year":"2006","journal-title":"Mol Cell Biol"},{"issue":"1","key":"pone.0250974.ref009","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1210\/en.2007-0734","article-title":"Targeted deletion of a distant transcriptional enhancer of the receptor activator of nuclear factor-kappaB ligand gene reduces bone remodeling and increases bone mass","volume":"149","author":"C Galli","year":"2008","journal-title":"Endocrinology"},{"issue":"2","key":"pone.0250974.ref010","doi-asserted-by":"crossref","first-page":"416","DOI":"10.1002\/jbmr.2698","article-title":"St. John HC, Danielson AL, and Pike JW. 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